Genetic and Molecular Characterization of a New EMS-Induced Mutant without the Third Glucose Moiety at the C-3 Sugar Chain of Saponin in Glycine max (L.) Merr.

Abstract

Saponin, a secondary metabolite, is produced by various plant species, including soybean (Glycine max (L.) Merr.). Soybeans synthesize triterpenoid saponins, which are classified by their aglycone structure and sugar chain composition. Here, we characterized an ethyl methanesulfonate-induced mutant, PE1539, without saponin and with a glucose moiety at the third position of the C-3 sugar chain. The saponin phenotype of PE1539 is described by the accumulation of Ab-γg saponin and deficiency of Ab-αg saponin and DDMP-αg saponin, similar to a previously reported sg-3 mutant in soybean. Genetic analysis showed that the saponin phenotype of PE1539 is controlled by a recessive mutation. We mapped the gene responsible for the phenotype of PE1539 and the mapped region included Sg-3 (Glyma.10G104700). Further analysis of Sg-3 in PE1539 using DNA sequencing revealed a single-nucleotide substitution in the exon (G804A), resulting in a premature stop codon; thus, PE1539 produced a PSPG box-truncated protein. Saponin phenotype analysis of the F2 population—from a cross between wild-type Uram and PE1539—showed that the phenotype of saponin was cosegregated with the genotype of Sg-3. Quantitative real-time PCR showed reduced expression of Sg-3 in PE1539 cells. Together, our data indicate that the saponin phenotype of PE1539 results from a mutation in Sg-3.